眼科研究 2000年第3期第18卷 实验研究
作者:张晓红 张红 汤欣 季建 孙慧敏
单位:300070天津医科大学世界人工晶体中国天津培训中心
关键词:白内障;晶状体囊膜;成分
摘要 目的 研究白内障患者晶状体囊膜中的成分。方法 采用间接免疫荧光法研究白内障术中取出的患者晶状体囊膜的成分。结果 Ⅳ型胶原、层粘蛋白阳性率为100%,部分患者晶状体囊膜中有纤维连接蛋白、Ⅰ型胶原存在。有些囊膜同时具有上述4种成分。结论 白内障患者的晶状体囊膜主要成分依然为Ⅳ型胶原和层粘蛋白,但也会有纤维连接蛋白、Ⅰ型胶原等成分存在。这些细胞外基质成分可能在白内障术后晶状体上皮细胞的移行、增殖中发挥作用。
分类号 R776
The components of cataract lens capsule
Zhang Xiaohong,Zhang Hong,Tang Xin
(International Intraocular Implant Training Center,Tianjin Medical University,Tianjin 300070)
Abstract Objective Posterior capsule opacification (PCO) is a common complication after cataract surgery.Several previous studies suggest that the residual lens epithelial cells (LECs) proliferation,migration and differentiation onto the posterior capsule lead to PCO.The aim of this study was to detect the extracellular matrix components in cataract lens capsule,in order to find the biological basis for LECs’ action.Methods The lens capsules obtained during cataract surgeries were pasted on the glasses coated with 3′-aminopropyl triethoxy-silane(APES).After they were dried at room temperature,they were assayed by indirect immunofluorescence to detect the presence of type Ⅰ and Ⅳ collagen,laminin,fibronectin.The secondary antibody was labeled by FITC.Results All the capsules examined were Ⅳ collagen and laminin positive.While only part of the capsules showed positive reactions with anti-fibronectin antibody or anti-type I collagen antibody.The positive percentages of fibronectin,type I collagen were 62.8% and 91.4%,respectively.It was noticeable that some capsules had the presence of all four extracellular matrix components.Conclusion Compared to the normal lens capsule,the main components of the cataract counterpart are still type Ⅳ collagen and laminin.But fibronectin and type I collagen are indeed present in some patients’ capsules.These extracellular matrixes may play a role in the migration and proliferation of lens epithelial cells after cataract surgery.
Key words cataract lens capsule components
白内障是目前居首位的致盲性眼病,虽然人工晶状体植入术可使患者恢复视力,但术后后囊混浊的发生却大大降低了手术的效果。晶状体最外层为晶状体囊,正常的晶状体囊中含有Ⅳ型胶原(type Ⅳ collagen)、层粘蛋白(laminin)等成分,在胚胎期还有纤维连接蛋白(fibronectin)[1]。目前的研究认为,后囊混浊的发生与白内障术后残留的晶状体上皮细胞的移行、增殖、化生有关。本文采用免疫组织化学方法,研究白内障患者晶状体囊膜的成分,以期发现晶状体上皮细胞移行、增殖的生物学基础。
1 材料与方法
1.1 人工晶状体植入术中取白内障患者晶状体前囊膜80例,其中男44例,女36例。年龄最大85岁,最小19岁。其中先天性白内障5例,外伤性白内障2例,白内障性质待查1例,老年性白内障72例。将上述患者分为4组,每组20例,分别测定囊膜中是否有Ⅰ型和Ⅳ型胶原、层粘蛋白、纤维连接蛋白的存在。同一年龄段的患者尽可能分在不同组。各组平均年龄见表1。
1.2 在此基础上,术中另外取白内障患者晶状体前囊膜15例,其中男10例,女5例。年龄最大85岁,最小5岁,平均年龄56.9岁。将每一患者的囊膜分为4份,分别用来测定Ⅰ型和Ⅳ型胶原、层粘蛋白、纤维连接蛋白的存在。本组患者中先天性白内障2例,外伤性白内障1例,老年性白内障12例。
1.3 采用间接免疫荧光法测定上述囊膜中的成分。首先将术中取下的囊膜贴于涂有防脱片剂APES的玻片上,晾干后固定囊膜,再经羊血清封闭后,加入一抗孵育60min(其中鼠抗人Ⅳ型胶原单抗购自美国Zymed公司,鼠抗人纤维连接蛋白单抗由英国Novocastra公司购买,鼠抗人Ⅰ型胶原单抗和兔抗人层粘蛋白抗体购自美国Sigma公司)。囊膜经PBS液漂洗后,与FITC标记的二抗孵育30min。再经PBS液漂洗、封片,于荧光显微镜下观察。囊膜上有黄绿色荧光者为阳性。每次实验均设阴性对照。
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